Objective To evaluate the potential of specific mRNA marker keratin 19(K19) to detect micrometastasis by reverse transcriptase polymerase chain reaction (RT-PCR) .Methods One hundred and ninty four regional lymph nodes harvested from 6 cases of benign diseases, 4 cases of breast carcinoma, 5 cases of gastric carcinoma and 12 cases of colorectal carcinoma patients were examined by conventional pathology and amplifying tissue specific K19 mRNA by RT-PCR separately, then the two methods were compared with each other. Results None of the 34 lymph nodes which were pathological metastasis-negative from benign diseases expressed K19 mRNA by RT-PCR, all of the 28 regional lymph nodes which were pathological metastasis-positive from malignant cases showed trains of K19 mRNA by RT-PCR. Of the 132 lymph nodes which were pathological metastasis-negative from malignant cases, 11 lymph nodes were detected with micrometastasis by genetic diagnosis.Conclusion Genetic diagnosis of lymph node micrometastasis is more sensitive than conventional pathology and has diagnostic value and merits further study.
ObjectiveTo explore the variation of the structure of the intestinal flora between healthy people and patients with obstructive jaundice perioperatively. MethodsFrom February 2013 to August 2014, 20 patients with obstructive jaundice and 10 healthy persons (normal control group) in our hospitol were selected as the research object. The first stool specimens of the research object after admission were obtained and the total fecal bacteria DNA were extracted. After polymerase chain reaction amplification, the changes in the structure of bacterial flora were dynamic observed by using denaturing gradient gel electrophoresis (DGGE), and the gel bands were analyzed by using Quantity One software. The similarity and diversity of flora structure, and principal component analysis (PCA) were analyzed. ResultsSignificant differences of colonic microflora were found between patients with obstructive jaundice and healthy people; advantage intestinal flora in obstructive jaundice patients was significant lower than the normal control group. With the extension of time and degree of obstruction aggravated, a descending trend was found in number, abundance, and diversity of the intestinal microflora (P < 0.05). ConclusionThere is significant differences in the structure of colon bacteria in patients with obstructive jaundice and healthy persons.
Objectives To study the relationships between methylenetetrahydrofolate reductase(MTHFR ) gene polymorphism and diabetic retinopathy (DR) in Chinese Han race． Methods With polymerase chain reaction and restriction fragment length polymorphism (PCR-FLP), MTHFR gene 677 T mutation (cytosine is replaced by thymine in No. 677 site) was detected in 85 health controls, 62 with DR and 117 without DR of type 2 diabetics comfrimed by ophthalmoscope． Results The frequency of MTHFR variant genotypes and alleles of DR in Chinese Han race．patients were signigicantly higher than those without retinopathy and healthy controls (Plt;0．01)． Conclusions The results suggested that MTHFRgene C677T mutation was probably one of the genetic risk factors of diabetic retinopathy in Chinese Han rase． (Chin J Ocul Fundus Dis, 2001,17:198-200
Abstract： The first generation scaffolds of bare metal stents （BMS） and the second generation of drug eluting stents （DES） have been widely used in the treatment of coronary heart diseases. However, long term incidences of major adverse cardiovascular events and revascularization treatments are still high because of in-stent re-stenosis and thrombosis. These may be caused by chronic inflammations and vascular wall damages due to persistent metal stents stimulation. What’s more, the eluting drugs within metal stents could also disturb normal growth of vascular endothelial cell, intima, tunica media, smooth muscle and epimysium. Therefore, in order to meet these demands several fully biodegradable scaffolds and drug carried stents have been manufactured using polymers polyester, polycarbonate and polyphosphate, etc. Among them, the security and histo-and hemo-compatibilities of coronary scaffolds made from poly-lactic acid （PLA）, poly-glycolic acid（PGA）, chitosan as coating, poly-caprolactone （PCL） and other copolymer like poly-lactic-co-glycolic acid （PLGA） have been testified to be sound. Nevertheless, there exist several different shortages for these stents such as tensile strength deficiency and slow degradation. PLA is hard and brittle with slow degradation, while PGA is soft with insufficient support force and fast degradation. Whether stents degrade too fast or too slow, they could not supply sufficient strength and effective support after implantation, and also they may cause target vascular injuries and elastic shrink inducing restenosis and thrombosis in long terms. Using optimized molar ratio component of PLA and PGA with chitosan coating, we can get sound composite materials with better biocompatibility, moderate degradation （approximately 3 - 6 months of completedegradation）, adequate mechanical strength, lower inflammatory response and good range of extension, and establish an experiment ground for fully biodegradable vascular scaffolds fabrication.
Objective To validate the different expressions of human fxyd6 gene between normal bile duct tissues and malignant tumor tissues, and to observe the subcellular localization of human fxyd6 gene in human cholangiocarcinoma cells. MethodsThe different expressions between normal bile duct tissues and malignant tumor tissues were identified by RT-PCR. In situ polymerase chain reaction （IS-RT-PCR） was applied to detect the subcellular localization of fxyd6 gene in paraffin sections of human cholangiocarcinoma cells. Image analysis software was used to semiquantitatively determine the difference between normal and malignant tissues. ResultsHuman fxyd6 gene was highly expressed in cholangiocarcinoma tissues and lowly expressed in normal ones. There was a significant difference between the expressions of carcinoma cells and normal cells （P＜0.05）. IS-RT-PCR showed that fxyd6 gene localized in the kytoplasma of epithelial cells of human cholangiocarcinoma. ConclusionHuman fxyd6 gene may act as an essential component of the malignant transformation process in human cholangiocarcinoma.
Objective To probe the association between HLA-B27 subtype and acute anterior uveitis(AAU). Methods The HLA-B27 gene of 49 AAu patients were inspected by means of polymerase chain reaction(PCR), and the B27 subtypes of AAU patients were analyzed by means of DNA sequencing and simulated nucleotide hybridization in computer. Results Twenty-nine(59.39%)of 49 AAU patients were B27 positive, and among them 13 (44.00%)were B2704 and 16(56.00%)B27052 in subtypes．No statistically significant difference in most clinical features were found between the patients with B2704 and B27052.In B27 pasitive patients,8 (seven is B27052 and one is B2704) of them were complicatde with ankylosing spondylitis(AS). Conclusions The AAU patients with either B2704 or B27052 have similar clinical feature.The B27052 gene may be an important factor for complicated AS in AAU patients. (Chin J Ocul Fundus Dis, 1999, 15: 139-142)
Objective To analyze the association of human leucocyte antigen (HLA)-DRB and -DQB alleles with Ealesprime; disease, and to investigate the potential immunogenetics mechanism of Ealesprime; disease. Methods Gene loci of HLA-DRB and -DQB1 alleles were detected by polymerase chain reaction-sequence specific primer (PCR-SSP) in 27 Han-nationality patients with Ealesprime; disease in Northern China and 30 age and sex-matched normal persons as control, then statistics package for social science (SPSS) for Windows ver 13.0 software was used to analyze the distribution features of frequency of HLA-DRB and -DQB1 alleles in the two groups. Results Compared with the control group, the frequency of HLA-DRB104 allele increased obviously in the patients with Ealesprime; disease［odds ratio (OR)=3.20 ，OR 95% confidence interval（CI）=1.00-10.21, and P=0.047］. Simultaneously, no statistically significant difference of the distribution of any other DRB or DQB1 allele between the two groups was found (Pgt;0.05). Conclusions In hannationality people in Northern China, DRB104 is positively associated with Ealesprime; disease, suggesting that DRB104 may confer a major influence on Ealesprime; disease. Turbulence of immune function begotten by infect-agents attack may occur in the individuals with Ealesprime; disease due to the specific hereditary diathesis of HLA, which may cause the occurrence and development of Eales disease. (Chin J Ocul Fundus Dis, 2006, 22: 90-93)
ObjectiveTo investigate the expressions and clinical significance of human telomerase reverse transcriptase (hTERT) mRNA and γglutamyl transpeptidase mRNA-H (GGT mRNA-H) in the peripheral blood of small hepatocellular carcinoma (HCC) patients. MethodsThe expressions of hTERT mRNA and GGT mRNA-H were detected in the peripheral blood of thirty patients with small HCC by RT-PCR, eighteen patients with benign liver diseases, and twelve normal volunteers. ResultsThe positive rate of hTERT mRNA and GGT mRNA-H expression in patients with small HCC were 80.0% (24/30) and 46.7%(14/30), respectively. In patients with hepatitic cirrhosis the positive rate of hTERT mRNA expression was 33.3% (6/18), while the expression of GGT mRNA was not detected. Both the expressions of hTERT mRNA and GGT mRNA-H were negative in all normal volunteers. The combination analysis of hTERT mRNA and GGT mRNA-H expression achieved positive rate of 86.7% in the diagnosis of small HCC, which was significantly higher than the positive rate of AFP (26.7%)， Plt;0.05. ConclusionThe hTERT mRNA and GGT mRNA-H are significantly expressed in small HCC patients, the combination analysis of hTERT mRNA and GGT mRNA-H seems to be useful in the early diagnosis of small HCC.
Objective To investigate the role of β-catenin gene in breast tumorigenesis by detecting mutation and expression of β-catenin gene in breast hyperplasia and breast cancer. Methods Mutation and expression of β-catenin gene in 42 breast cancer, 15 simple hyperplasia and 15 atypical hyperplasia were detected by polymerase chain reaction-single strand conformation polymorphism and immunohistochemistry. Results Normal expression of β-catenin occurred in tissue of breast simple hyperplasia. The rate of abnormal expression of β-catenin in tissue of breast atypical hyperplasia and breast cancer were 26.7% （4/15） and 59.5% （25/42）, respectively, which were higher than that of simple hyherplasia tissue (P＜0.05). And there was a markedly difference between the atypical hyperplasia tissue and breast cancer tissue (P＜0.05). Mutation of β-catenin gene wasn’t detected in this three kinds of tissues. Conclusion Abnormal expression of β-catenin plays an important role in human breast tumorigenesis, reason of abnormal expression of β-catenin isn’t mutation of β-catenin gene. Expression of β-catenin can be regulated by other mechanisms.
Objective To investigate the polymorphism of the vitamin D receptor gene (VDR)TaqⅠin relation to diabetic retinopathy. Method Fragment length discrepant allele specific PCR(FLDAS-PCR) were used to determine VDR genetypes in 158 patients with diabetic retinopathy and in 198 normal subjects. Results The frequency distribution of VDR genotypes in diabetic retinopathy patients was 106 (67.1%) in TT, 33(20.9%) in Tt, 19(12.0%) in tt; and in normal persons was 165 (83.3%) in TT, 23(11.6%) in Tt, 10 (5.1%) in tt. There was a significant difference between diabetic retinopathy patients and normal persons in distribution of VDR gene TaqⅠgenotypes(Plt;0.05). Conclusions There is some distribution alterations of VDR gene polymorphism in diabetic retinopathy patients. (Chin J Ocul Fundus Dis, 2006, 22: 94-96)